CN102576406A - Compact automated cell counter - Google Patents
Compact automated cell counter Download PDFInfo
- Publication number
- CN102576406A CN102576406A CN2010800457267A CN201080045726A CN102576406A CN 102576406 A CN102576406 A CN 102576406A CN 2010800457267 A CN2010800457267 A CN 2010800457267A CN 201080045726 A CN201080045726 A CN 201080045726A CN 102576406 A CN102576406 A CN 102576406A
- Authority
- CN
- China
- Prior art keywords
- cell
- sensor
- digital imagery
- image
- digital
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
- C12Q1/06—Quantitative determination
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Electro-optical investigation, e.g. flow cytometers
- G01N15/1434—Electro-optical investigation, e.g. flow cytometers using an analyser being characterised by its optical arrangement
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/0004—Microscopes specially adapted for specific applications
- G02B21/0008—Microscopes having a simple construction, e.g. portable microscopes
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/34—Microscope slides, e.g. mounting specimens on microscope slides
Abstract
Biological cells in a liquid suspension are counted in an automated cell counter that focuses an image of the suspension on a digital imaging sensor that contains at least 4,000,000 pixels each having an area of 2 x 2 [mu]m or less and that images a field of view of at least 3 mm<2>. The sensor enables the counter to compress the optical components into an optical path of less than 20 cm in height when arranged vertically with no changes in direction of the optical path as a whole, and the entire instrument has a footprint of less than 300 cm<2>. Activation of the light source, automated focusing of the sensor image, and digital cell counting are all initiated by the simple insertion of the sample holder into the instrument. The suspension is placed in a sample chamber in the form of a slide that is shaped to ensure proper orientation of the slide in the cell counter.
Description
The cross reference of related application
The application requires the U.S. Provisional Patent Application No.61/238 of submission on August 31st, 2009,534 right of priority, and its content is incorporated into this by reference.
Technical field
The field that the present invention relates to blood clotting and generally be used for the system that biological cell that convection cell suspends counts.The automatic cytological number system that focuses on of the present invention.
Background technology
In various clinical and research process; Cell count is interesting; Comprising leucocyte and erythrocytic counting, and this is of great value in the diagnostic procedure of various diseases or unusual condition and in the patient's who monitors the treatment of experiencing this disease or situation process.Can carry out artificial counting through the processes pair cell: known diluted sample thing is placed between the plate of optical clarity, these plates each other fully near (usually, be magnitude at interval) at 100 microns so that these cells form single layer; One zone of this layer is amplified to known multiple on specified dimension; And the cell in the zone that is exaggerated is counted through microscope.The artificial cell counter generally includes the grid that is engraved in the counting region to reduce user's burden.Qiu, the U.S. Pat 7,329 of J, 537B2 (on February 12nd, 2008 authorized, and was entitled as " Micro-Pattern Embedded Plastic Optical Film Device for Cell-Based Assays ") has described this grid and use thereof.No matter how this realizes that the artificial cell counting all is dull, and the user is easy to make mistakes.High dilution thing through using this sample usually helps counting to reduce the cell number in the counting region, and still, that part of cell that is counted often reduces to some extent, and the accuracy of this counting will descend.
Through using digital imaging system, made the robotization of cell count process become possibility.The example of this system is ImageJ; A kind of image processing program based on Java in the exploitation of national health institute; Collins, the article of T.J. " ImageJ for microscopy " (BioTechniques 43 (1Suppl.): 25-30, in July, 2007) carried out report to this.Gering; T.E. in article " A rapid method for counting nucleated erythrocytes on stained blood smears by digital image analysis " (J.Parasitol.90 (4): 879-81,2004), reported the situation of in hematological system, using ImageJ with C.Atkinson.Following document has further described automatic cytological counting: Chang, people's such as J.K. U.S. Pat 7,411, and 680B2,2008, August 12 authorized, and was entitled as " Device for Counting Micro Particles "; And Chang, people's such as J.K US patent application publication US 2006/0223165A1 announced, and was entitled as " Device for Counting Cells and Method for Manufacturing the Same " on October 5th, 2006.
The automatic cytological number system self comprises intrinsic statistical uncertainty, and this is to be caused by so-called " sampling error " usually, and this " sampling error " is meant error intrinsic when selecting to carry out automated enumeration regional.One of many limitations of present commercially available automatic cytological counter are: because the restriction of the optics in the instrument, the zone that pair cell is counted is compared with the occupied whole zone of sample has limited size.Because this has correspondingly limited the number of cell; And the number of the cell that is counted often reduces to some extent; Error will increase, thus the typical instrument of prior art be configured to have very long light path or very big areal coverage (this instrument on testing table occupied surface area) or both to realize acceptable accuracy.This has brought inconvenience to the user, particularly when this instrument will be used in the cellular incubation lid.
Summary of the invention
This paper has disclosed a kind of Cytometric fully independently instrument that is used for pin-point accuracy, and user intervention is minimum, and areal coverage is relatively very little, and highly also limited.Cell suspending liquid is placed in the consumable sampling receptacle, and the size of this container and dimension can vary widely, one of this container easily example be its external dimensions container similar with microslide.This container thus can be aspect structure and size the container described in the above-mentioned US 2006/0223165A1; This container has at least one flat and shallow internal chamber; The top of this internal chamber and bottom are the boundary with window that put down, optical clarity, and these windows can be plastic sheets and get enough near at interval so that layer of most of sample cell formation is that a cell is so dark.In this container, can comprise suitable entrance and exit, to allow filling up this chamber easily and fully with sample.Then, this container is put in this instrument, this container and linear optical path intersect in this instrument.One of employed in this article term " linearity " expression is in the path that does not have turning or other variation on the beam direction except turning that lens caused or changing.This container gets into this instrument through the slit at the specified altitude assignment place in this light path; And as hereinafter institute in more detail as described in; Instrument among some embodiment of the present invention comprises a plurality of characteristic items, is used for automatically regulating the height of sample so that focus on sample image.Some embodiment comprises a plurality of characteristic items, is used for making all instrumental functions when sampling receptacle is inserted this instrument, just to start working.
Description of drawings
Fig. 1 is the skeleton view of cell count instrument of the realization example of representative notion disclosed herein.
Fig. 2 is the figure of optics of the instrument of Fig. 1.
Fig. 3 is the skeleton view of the optics in the inside of instrument of Fig. 1.
Fig. 4 is the employed decomposition diagram that is used to constitute two plates of specimen slide in the instrument of Fig. 1.
Fig. 5 A is the figure of upper surface of top board of the specimen slide of Fig. 4.Fig. 5 B is the figure of lower surface of top board of the specimen slide of Fig. 4.
Embodiment
Between top optical window and bottom optical window; Cell suspending liquid is remained within the sampling receptacle; This top optical window and bottom optical window lean on enough closely; Make that the suspending liquid kept is a film, the transverse dimensions of this film is that its length of exposing and width are at least than its big one magnitude of thickness.Partly as a visual field, this visual field is projected on the digital imagery sensor for the whole zone of exposing (being transverse dimensions) of sample chamber or its transverse dimensions, and this sensor comprises at least about 4,000, and 000
(4,000,000) individual pixel or comprise approximately 4,000,000 to 10,000 in certain embodiments, 000 pixel, the size of each pixel are not more than about 2 * 2 μ m (4 μ m
2) or be about 0.5 * 0.5 μ m (0.25 μ m in certain embodiments
2) to 2 * 2 μ m (4 μ m
2) and in some embodiment of the latter, be about 1 * 1 μ m (1 μ m
2) to 2 * 2 μ m (4 μ m
2).Visual field by the sensor imaging is at least about 3 square millimeters, is about the 3-10 square millimeter usually.Complementary metal oxide semiconductor (CMOS) (CMOS) is an example that is used for the digital imagery sensor of this purpose.The example that meets the cmos sensor of these parameters is OV5620 and the OV5632 colour imaging device that can buy from the OmniVision company in California, USA Santa Clara city.Other example then can buy from the Aptina Imaging department of the Micron Technology company in California, USA San Jose city.Also can use the colorful digital imaging sensor.Through known digital counting method, such as mentioned above those, can realize that Flame Image Process is to count the cell in the image that cmos sensor was produced.
Along light path, can amplify the image of sample chamber by an enlargement factor, this enlargement factor is normally in about scope of 1.5 to 6 or in about scope of 1.5 to 3, and as an example, this enlargement factor is about 2.This can realize through a kind of double lens formula achromat assembly.The example of this lens subassembly is: near the lens of the 35mm focal length of sample, and near the lens of the 60mm focal length of sensor, and the aperture between these two lens.Thus, in this example, be 35mm near the lens of sample and the distance between the sample self, and be 60mm near the lens of sensor and the distance between the imager self.The enlargement factor of system is the ratio of the focal length of these two lens, is exactly 60mm/35mm=1.7 in this case.For example, the diameter of these two lens can be 12.5mm, and the diameter in above-mentioned aperture can be 6mm.Meeting produces other diameter of identical or roughly the same result and the lens of focal length are obvious to those skilled in the art.The areal coverage of this instrument is defined as that bigger in this instrument and the supporting base thereof zone of on the plane perpendicular to light path, being throwed.As stated, this instrument can be configured to have very little areal coverage, and particularly its area is less than 300 square centimeters areal coverage.
When using flat digital imagery sensor, negative lens can be positioned at this sensor below to block light signal at once and to proofread and correct the right focousing field curvature of this achromat.This field curvature is general in multiple optical system, also is called as Petzval curvature.In the embodiment of example, used 6mm diameter lenses with negative 18mm focal length.Lens thickness can change to some extent, can proofread and correct above-mentioned curvature and does not significantly reduce the visual field but preferably be selected to.
Through using conventional light source and the collimation lens between this light source and sample, can realize the illumination of this sample at the instrument base place.Use these parts, just can shine this sample, and need not diffusing globe through transillumination.Preferable light source is single white light-emitting diode (LED) with fluoresent coating.The example of this parts is: LUXEON
Rebel White; Parts number is LXML-PWN1-0050, can buy from the Philips Lumileds Lighting company in California, USA San Jose city.The example of collimation lens is that diameter is that 9mm and focal length are the lens of 18mm.The size of using these sizes and preceding text to mention can construct a kind of instrument, and the achromat of this instrument is to the 35mm about sample top greatly, the sensor of this instrument greatly about this achromat to top 60mm.Under the situation of achromat to thickness with about 13mm, the total distance between sample and the sensor may diminish to 108mm.Usually, the height of the light path of this instrument (that is, being defined as the arranging of a plurality of parts that extends to CMOS or other digital imagery sensor from light source in this article) can be 20cm or point more.In the preferable instrument within scope of the present invention; Use the counting of biddability a plurality of opticses to be installed to enclosure by a kind of mode of floating; Optical system is destroyed or lacks of proper care when shaking this instrument to avoid, such as when instrument generation decline or improper use or with another instrument or change when bumping against and this destruction or imbalance possibly occur.
As stated; Through a slit; Sampling receptacle (is considered that its size and shape is similar in appearance to microslide; To be called as specimen slide hereinafter) be received in the above-mentioned instrument, this slit is positioned at the position along above-mentioned light path, and this position equals the focal length of these lens from the distance of the right immediate lens of above-mentioned achromat.In its preferred embodiment; The whole height of this instrument is 30cm or littler, and can allow this instrument to be configured to have sufficiently high slit to allow the user cosily with this slit of hand insertion (promptly letting user's hand can not meet the desk at this instrument place) by use digital imagery sensor mentioned above (this sensor uses a large amount of small-sized pixel).Thus, this slit can have 60mm or bigger from the base of this instrument, preferably from this base 70-80mm is arranged.
In preferred embodiment of the present invention, through after inserting microslide, automatically regulating the height of this microslide, above-mentioned instrument provides the automatic focus of sample image.A kind of automatic focus means comprise uses the image processor chip, and this chip provides in the output from the image comparison within the array in a plurality of zones on the image of sensor.The example of this chip is can be from the Keil of Texas, USA Plano city ARM company
TMBought " Freescale Semiconductor MC9328MX21 "; Other example is clearly to those skilled in the art.The antipode sum of adjacent green pixel can be used as the image comparison value in the specific region of sensor array, and when this image comparison value reaches maximum, has just realized optimum focusing.Through being connected to the above-mentioned gear motor of taking in the microslide supporting plate in the slit, can regulate above-mentioned focusing, that is, motor will make the microslide supporting plate move up or down to change the focusing situation of image when rotating.This correlative value is detected in each position at motor, then, turns back to the position of high correlative value of generation.In many embodiment of this instrument, this automatic focus can occur in 15 seconds or less time in.
The annex that can be this instrument provides is the standard microslide, is used for quality control, such as the ability that is used to verify that accuracy that living cells and dead cell are counted and this instrument appropriately focus on.This standard microslide can have the external dimensions identical with specimen slide; But it is different with sample chamber; This standard microslide can have stamp dark spots and ring above that; These spots are simulated dead cells and in the digital imagery sensor, are taken as dead cell and detect, and these rings are then simulated living cells and in the digital imagery sensor, are taken as living cells and detect.
In some embodiment of many notions described herein,, just started the performed function (comprising automatic focus and cell count) of this instrument through inserting specimen slide simply.Through placing a non-contacting optical reflection sensor on the microslide supporting plate in above-mentioned slit or in the above-mentioned slit, just can realize this startup.The example of right sensors is a kind of like this sensor, and it can be launched infrared beam and detect the object in about 1 millimeter of sensor aperture through the reflected signal that detects this light beam.When microslide was inserted into, it is maximum that this reflected signal will reach, and high signal will start automatic focus and cell count mechanism.An example that can be used for the sensor of this purpose is " the reflective object sensor of QRE1113 ", and this can buy from the Fairchild Semiconductor in California, USA San Jose city.Other example is clearly to those skilled in the art.
In the instrument of practical implementation many characteristics described herein, another characteristic that can comprise is: detect the cell in the sample of being infected with by vital stain automatically.Vital stain is the coloring agent that is used for preferentially being infected with dead cell, and the difference between cell of being infected with by this coloring agent and the cell be not infected with is to realize through the pixel of using different colours.Trypan blue is an example of vital stain, and eosin and propylidene are other examples.Trypan blue makes the blue light transmission and makes the ruddiness decay, and through the blue pixel in the movement images sensor and the intensity of red pixel, this instrument just can determine whether to exist the cell of being infected with by vital stain.Other dyestuff will provide similar color differentiating, as long as suitable to dyestuff self.Those that the picture processing chip that comprises this automatic detected characteristics comprises that preceding text are mentioned and be easy to buy.Can programme to this instrument, with through on two or more planes enterprising line focusing eliminate any possible living cells error of omission, living cells detects and can reach extra high accuracy thus.Through using optical filter to control the illumination bandwidth, perhaps, just can further increase the contrast between living cells and the dead cell through selecting the very narrow light source of spectrum (such as the LED of particular color but not white light).For example, can use 585nm optical filter, so that above-mentioned illumination is matched with the peak absorbtivity wavelength (its peak absorbance is 586nm) of trypan blue dyestuff with about 20nm bandwidth.When shining this sample through this optical filter, it is darker that dead cell will seem.
In the preferable instrument within the scope of the invention, the contributive all functions of Cytometric acquisition in the sample all are comprised in this tool housing, thus, can realize the complete operation of this instrument and need not to use external mechanical or computing machine.In these functions, comprised following: the height through changing specimen slide comes automatic focus, finding best focal plane, thereby cell and background is made a distinction; Confirm whether this sample is infected with by trypan blue or other vital stain; When detecting vital stain, carry out many focal planes and analyze, make and on a plurality of focal planes, write down each cell, to prevent the error of omission of living cells; Integrated dilution counter is used for confirming the volume of the cell suspending liquid that will use; When the user selects, can on display, produce the visual image of these cells and can amplify so that carefully check these cells; And when the user selects, can these results be outputed to USB flash memory driver or thermal printer or other external printer.Through the described non-contacting optical reflection sensor of preceding text, to insert specimen slide simply and just can start all these functions, and in many cases, the execution of these functions is accomplished in 30 seconds or less time.
The instrument that accompanying drawing is described has comprised many above-mentioned characteristics, and is used as the realization example of many functions described herein.
Fig. 1 has described a kind of automatic cell counter instrument 11 that is in its stand up position, its son that comes to this when on testing table, using.The visible part of this instrument is: shell 12, base for supporting 13, display screen 14, control panel 15, and the slit 16 that is used to insert specimen slide 17.Display screen shows the process that cell count is analyzed, and has identified the various functions that just are being performed of this instrument, and to the user multiple option is provided, and is used for multiple function and is used for being illustrated in the image of the cell of specimen slide.
Fig. 2 has described the various parts of light path of the instrument internal of Fig. 1, and specimen slide 17 has been placed to this light path and has suffered.Specimen slide 17 is levels, and is positioned at the top as the led board 22 of light source.Collimation lens 23 makes the light from this LED near this specimen slide the time, become parallel.Achromat to 24 between specimen slide 17 and sensor 25.These right two lens 26,27 of achromat were left by an aperture in 28 minutes.Smooth lens 29 are adjacent below sensor 25.
Fig. 3 has described main optical module, shows: microslide supporting plate 31, and specimen slide 17 is partly inserted wherein; Led board 22; Illumination (collimation) lens 23; Gear motor 32 is used to regulate the microslide height to focus on this image; And imaging len pipe 33, be terminated at assembly parts 34 to take in the cmos sensor plate.Also show main printed circuit board (PCB) 35 among the figure, be used to control the various functions of this instrument and comprise that the motor driven chip is with control motor 32.Plate 35 is positioned within the shell, and its position with respect to optical module has been reflected in this plate position in the drawings.
Employed specimen slide in the instrument among the figure before Fig. 4,5A and 5B show.Fig. 4 is a skeleton view, and microslide 17 is to be made up of two flat boards 42,43 (but being shown as separately among the figure) that join to together.This microslide comprises two sample chamber, is labeled as " A " and " B " respectively, and these two sample chamber longitudinally are separated from each other and laterally depart from each other along this microslide.Constitute following dull and stereotyped 43 zone the 44, the 45th of sample chamber bottom surface, process by optically transparent material, as the respective regions of upper flat plate 42, these respective regions be located immediately at play on the flat board these regional above and constitute the end face of sample chamber.In the present embodiment; Following dull and stereotyped 43 is thicker in to microslide rigidity being provided than upper flat plate 42, thereby and the relatively thin top window that just can allow each sample chamber of upper flat plate 42 than bottom windows thinner and in fact thin as far as possible in cmos sensor the image of realization high order focusing.Thus, each sample chamber departs from from the central plane of microslide to some extent, and compares more near upper flat plate 42 with following dull and stereotyped 43.
Fig. 5 A and 5B are respectively the end face 51 of upper flat plate 42 and the planimetric map of bottom surface 52, and bottom surface 52 is to join down that surface of dull and stereotyped 43 to.Each sample chamber is defined by the depression in the bottom surface of upper flat plate 53 (Fig. 5 B), and this has further reduced to form at each sample chamber top the thickness in the zone of optical clarity window.In an example of the size of microslide, the thickness except the zone 53 of caving in the upper flat plate is 0.65mm, and following dull and stereotyped thickness is 1.00mm, and total slide thickness is 1.65mm.The degree of depth of depression 53 is 0.100mm, and having formed the degree of depth thus is the sample chamber of 0.100mm, and this is the standard model chamber depth of artificial hemacytometer.Each sample chamber has two inlets or exports 54,55, and two relative vertical ends of elongated chamber respectively have one.The microslide open-top overflow four corners that zone 56,57,58,59 is positioned at each sample chamber to hold unnecessary sample, guarantee that thus sample chamber filled up by sample rightly.
Because each sample chamber more but not down dull and stereotyped 43 near upper flat plate 42; So when microslide was inserted in the cell counter rightly and upper flat plate 42 is positioned at the top and therefore the thinnest optical window also is positioned at the top, this microslide is the best function of performance just.In order to ensure inserting this microslide by this orientation, the angle place relative at two diagonal line of microslide forms recess 61,62.The inside surface of the slit in the cell counter (microslide just is inserted in this slit to start the multiple function of this cell counter) comprises the contour feature complementary with these recesses.Thus, the profile of the complementation in these recesses and the slit can prevent to have turned upside down when the user from inserting microslide, promptly upper flat plate 42 in the bottom (should at the top).The symmetry arrangement of these recesses is also complementary with the symmetry arrangement of two sample chamber, and when the insertion microslide, allows arbitrary terminal elder generation to insert, and prevents simultaneously to fall contrary position insertion microslide (turning upside down).Because microslide is consumable article preferably, thus it can be used to the different moment to two independently sample carry out cell count and measure, in case and two chambers all have been used then this microslide can be dropped and no longer be used.
It will be apparent to those skilled in the art that under the situation of guaranteeing appropriate orientation, the structure of specimen slide can change to some extent.Thus, can change the arranging of recess, number and shape, the relative position of each other also can change as the number of sample chamber and in microslide.The material of structure also can broadly change, and can be that any material that can form the optical clarity window does not promptly react with sample and enough firm so that any material in the insertion cell counter.Gather the example that (methacrylate) and polycarbonate are spendable materials.Other material is conspicuous to those skilled in the art.Likewise, the joint of above-mentioned plate can be realized through conventional means.Laser bonding and ultra-sonic welded are examples.
In the claims by the appended claims herein, term " " or " one " are intended to expression " one or more ".When describing step or element, term " comprises " and waits its variant to be intended to represent that the interpolation of other step or element is optional and is not exclusive such as " included " and " comprise ".Whole patents that this instructions is quoted, patented claim and other disclosed reference material integral body by reference are incorporated into this.Any difference between the clear and definite teaching of any reference material that this paper quoted or any general prior art and this instructions is intended to advantageously be included in this instructions teaching.It comprises any inconsistent between the definition that this instructions clearly provided of the definition that admits in this field of speech or phrase and same words or phrase.
Claims (26)
1. automatic cell counter comprises:
Optics;
Be used to take in the sample splint of specimen slide; And
Be used to keep the shell of said optics and sample splint,
Said optics comprises:
Light source;
The digital imagery sensor; And
Lens, said lens are placed with guiding and pass said sample splint from the light of said light source and arrive said digital imagery sensor,
Said optics is arranged in the linear optical path; And be arranged to make when the specimen slide that comprises cell suspending liquid is installed in the said sample splint; The image of said cell suspending liquid is projected on the said digital imagery sensor; Said digital imagery sensor comprises at least about 4,000,000 pixel and to the 3mm at least at sample splint place
2The visual field be carried out to picture, the area of each pixel is 2 * 2 μ m or littler.
2. automatic cell counter as claimed in claim 1, wherein,
It is about 4,000,000 to 10,000 that said digital imagery sensor comprises, 000 pixel, and the area of each pixel is about 0.5 * 0.5 μ m to 2 * 2 μ m, and the field of imaging is about 3mm
2To 10mm
2
3. automatic cell counter as claimed in claim 1, wherein,
Said light path is vertical orientated, and said shell is installed to a supporting base, and said supporting base is the same with said shell at least wide, and the horizontal area that said shell and supporting base occupy separately is not more than 300cm
2, the overall height of said light path is not more than 20cm.
4. automatic cell counter as claimed in claim 1, wherein,
Said digital imagery sensor comprises the device of the correlative value between the adjacent area that is used to produce said image; Said sample splint has the adjustable height along said light path, and said cell counter also comprises and is used for regulating the device that said height is focused up to said image automatically in response to said correlative value.
5. automatic cell counter as claimed in claim 4 also comprises:
The digital imagery device is used for the cell of said visual field is counted.
6. automatic cell counter as claimed in claim 5, wherein,
When being inserted into specimen slide in the said sample splint, just automatically start the automatic adjusting and the digital imagery device of the said height of said sample splint.
7. automatic cell counter as claimed in claim 1, wherein,
Said digital imagery sensor is put down, and said lens comprise that achromat is right, and said achromat is to having the focousing field at said digital imagery sensor place, and said automatic cell counter also comprises the device of the curvature that is used to proofread and correct said focousing field.
8. automatic cell counter as claimed in claim 7, wherein,
The said device that is used to proofread and correct the curvature of said focousing field is and the adjacent negative lens of said digital imagery sensor.
9. automatic cell counter as claimed in claim 1, wherein,
Said light source is that spectrum is limited, and in said imaging sensor, to produce an image, said image is made differentiation between living cells and dead cell.
10. automatic cell counter as claimed in claim 1, wherein,
Said digital imagery sensor is the colorful digital sensor.
11. automatic cell counter as claimed in claim 3, wherein,
When said supporting base is positioned at surface last time, the height of said shell is that 30cm or littler and said sample splint are 6cm at least in said surface.
12. automatic cell counter as claimed in claim 1 also comprises:
Quality control microslide, said quality control microslide have and are used to be installed to the size on the said sample splint, and are printed on pattern on it with simulation living cells and dead cell.
13. the specimen slide that the cell that is used for pair cell suspending liquid is counted, said specimen slide comprises:
Flat board with internal chamber, said internal chamber is the boundary with the top window and the bottom windows of optical clarity; And
Orientation recess in said flat board, be used to guide said flat board by a kind of be orientated get into the cell count instrument make thus said top window at said instrument inner face towards a selected direction.
14. specimen slide as claimed in claim 13, wherein,
Said top window is thin more a lot of than said bottom windows.
15. specimen slide as claimed in claim 13 comprises:
Single flat board; In said single flat board, first and second internal chamber are arranged; Said internal chamber laterally is positioned relative to each other in said flat board, and each internal chamber all has the top window and the bottom windows of optical clarity, and said single flat board comprises the first and second orientation recesses; Be used to guide said flat board to get into said cell count instrument by orientation, the top window that makes optical clarity thus at said instrument inner face towards a selected direction.
16. the method that the cell that is used for pair cell suspending liquid is counted, said method comprises:
(a) aliquot with said cell suspending liquid is placed in the internal chamber of specimen slide, in said chamber, to form the film of said suspending liquid;
(b) through being inserted into said specimen slide on the sample splint and said sample splint being placed in the linear optical path that is formed by optics, on the digital imagery sensor, produce the 3mm at least in the said film
2The digital picture of visual field; Said digital imagery sensor comprises at least 4; 000,000 pixel, the area of each pixel are 2 * 2 μ m or littler; Said optics comprises light source, said digital imagery sensor and lens, and said lens are placed with guiding and pass said sample splint from the light of said light source and arrive said digital imagery sensor; And
(c) cell in the said digital picture is carried out digital counting.
17. method as claimed in claim 16, wherein,
It is about 4,000,000 to 10,000 that said digital imagery sensor comprises, 000 pixel, and the area of each pixel is about 0.5 * 0.5 μ m to 2 * 2 μ m, and the field of imaging is about 3mm
2To 10mm
2
18. method as claimed in claim 16, wherein, step (b) is through comprising that following multiple function carries out:
Through moving said sample splint, automatically focus on said digital picture;
Detect whether the vital staining agent is arranged in the said aliquot;
When detecting the vital staining agent, produce the digital picture of a plurality of focal planes in the internal chamber of said specimen slide; And
Processing on said digital imagery sensor formed image confirming the cell count in the said aliquot,
Above-mentioned multiple function is all carried out in above-mentioned shell.
19. method as claimed in claim 16, wherein,
Said light path is vertical orientated; And said sample splint and said optics are maintained in the shell, and said shell is installed to a supporting base; Said supporting base is the same with said shell at least wide, and the horizontal area that said shell and supporting base occupy separately is not more than 300cm
2, the overall height of said light path is not more than 20cm.
20. method as claimed in claim 16 also comprises: (a ') automatically produces the correlative value between the adjacent area of said image, and, in response to correlative value, said sample splint automatically is adjusted to a height to focus on said image along said light path.
21. method as claimed in claim 16, wherein,
When being inserted into said specimen slide in the said sample splint, setting up procedure (a ') and (b) automatically just.
22. method as claimed in claim 16, wherein,
Said digital imagery sensor is put down, said lens comprise achromat to and negative lens, said achromat is to having the focousing field at said digital imagery sensor place, said negative lens is oriented to proofread and correct the curvature of said focousing field.
23. method as claimed in claim 16, wherein,
Said light source is that spectrum is limited, and in said imaging sensor, to produce an image, said image is made differentiation between living cells and dead cell.
24. method as claimed in claim 16 also comprises:
Handle said cell suspending liquid with the vital staining agent, in said imaging sensor, produce an image thus, said image is made differentiation between living cells and dead cell.
25. method as claimed in claim 16, wherein,
Said digital imagery sensor is the colorful digital sensor.
26. method as claimed in claim 16, wherein,
Step (c) comprises said optics is focused on a plurality of planes in the said internal chamber.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US23853409P | 2009-08-31 | 2009-08-31 | |
| US61/238,534 | 2009-08-31 | ||
| PCT/US2010/047143 WO2011026029A1 (en) | 2009-08-31 | 2010-08-30 | Compact automated cell counter |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102576406A true CN102576406A (en) | 2012-07-11 |
| CN102576406B CN102576406B (en) | 2014-07-23 |
Family
ID=43628421
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201080045726.7A Active CN102576406B (en) | 2009-08-31 | 2010-08-30 | Compact automated cell counter |
Country Status (8)
| Country | Link |
|---|---|
| US (2) | US8570370B2 (en) |
| EP (1) | EP2473948B1 (en) |
| JP (1) | JP5682974B2 (en) |
| CN (1) | CN102576406B (en) |
| AU (1) | AU2010286499B2 (en) |
| CA (1) | CA2772376C (en) |
| SG (1) | SG178555A1 (en) |
| WO (1) | WO2011026029A1 (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105492887A (en) * | 2013-03-15 | 2016-04-13 | 理查德·哈理·特纳 | A system and methods for the in vitro detection of particles and soluble chemical entities in body fluids |
| CN106030343A (en) * | 2013-12-17 | 2016-10-12 | 阿兰蒂克微科学股份有限公司 | Dosimeters including lensless imaging systems |
| CN110914666A (en) * | 2017-05-19 | 2020-03-24 | 兴盛生物科技股份有限公司 | System and method for counting cells |
| CN114067315A (en) * | 2021-10-23 | 2022-02-18 | 广州市艾贝泰生物科技有限公司 | Cell counting method, cell counting device, computer device, and storage medium |
Families Citing this family (28)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8570370B2 (en) | 2009-08-31 | 2013-10-29 | Bio-Rad Laboratories, Inc. | Compact automated cell counter |
| US9075225B2 (en) | 2009-10-28 | 2015-07-07 | Alentic Microscience Inc. | Microscopy imaging |
| EP2494400B1 (en) | 2009-10-28 | 2021-12-08 | Alentic Microscience Inc. | Microscopy imaging |
| US8852524B2 (en) | 2010-01-12 | 2014-10-07 | Bio-Rad Laboratories, Inc. | Cell counting slide with lateral reservoir for promoting uniform cell distribution |
| US9001200B2 (en) | 2010-01-12 | 2015-04-07 | Bio-Rad Laboratories, Inc. | Cell characterization using multiple focus planes |
| EP2503376B1 (en) * | 2011-03-24 | 2020-02-19 | Fisba Ag | Device and method for imaging |
| US9354155B2 (en) | 2011-05-31 | 2016-05-31 | Bio-Rad Laboratories, Inc. | Cell counting systems and methods |
| US10502666B2 (en) | 2013-02-06 | 2019-12-10 | Alentic Microscience Inc. | Sample processing improvements for quantitative microscopy |
| CA2953620C (en) | 2013-06-26 | 2020-08-25 | Alentic Microscience Inc. | Sample processing improvements for microscopy |
| EP3100025B1 (en) | 2014-01-30 | 2021-03-03 | BD Kiestra B.V. | System and method for imaging biological samples disposed in culture media |
| WO2016013395A1 (en) | 2014-07-22 | 2016-01-28 | 株式会社日立ハイテクノロジーズ | Cell dispersion measurement mechanism, and cell subculture system utilizing same |
| JPWO2016017591A1 (en) * | 2014-08-01 | 2017-06-01 | シャープ株式会社 | Inspection instrument, inspection device, inspection kit, and measurement method |
| US10625259B1 (en) | 2014-11-26 | 2020-04-21 | Medica Corporation | Automated microscopic cell analysis |
| US11478789B2 (en) | 2014-11-26 | 2022-10-25 | Medica Corporation | Automated microscopic cell analysis |
| US20170328924A1 (en) | 2014-11-26 | 2017-11-16 | Ronald Jones | Automated microscopic cell analysis |
| CA3211036A1 (en) | 2015-04-23 | 2016-10-27 | Bd Kiestra B.V. | A method and system for automated microbial colony counting from streaked sample on plated media |
| JP6777726B2 (en) | 2015-04-23 | 2020-10-28 | ビーデー キーストラ ビー．ヴィー． | Colony contrast collection |
| CN108351282B (en) * | 2015-06-01 | 2020-10-27 | 耐克思乐生物科学有限责任公司 | Methods for cell counting and viability measurement |
| JP1552333S (en) * | 2015-06-23 | 2016-06-20 | ||
| JP6586300B2 (en) * | 2015-06-23 | 2019-10-02 | オリンパス株式会社 | Cell counting device |
| JP1567588S (en) * | 2015-11-05 | 2017-01-23 | ||
| WO2018107105A1 (en) * | 2016-12-08 | 2018-06-14 | Essenlix Corporation | Assay sample cards and adaptors and use of the same |
| US11047845B1 (en) | 2017-11-15 | 2021-06-29 | Medica Corporation | Control material and methods for cell analyzers |
| CN109061214B (en) * | 2018-10-31 | 2023-12-19 | 江苏卓微生物科技有限公司 | Porous sample injection device |
| CN109825427A (en) * | 2019-02-25 | 2019-05-31 | 广州牛顿光学研究院有限公司 | A kind of full-automatic cell activity analysis system |
| BG67480B1 (en) | 2019-10-30 | 2022-12-15 | "Милкотроник" Оод | Device for differential counting of microparticles in biological liquids |
| TWI740420B (en) | 2020-03-19 | 2021-09-21 | 邦睿生技股份有限公司 | Validation method for testing biological sample quality confirmation test piece and testing biological sample quality detector |
| CN116046647B (en) * | 2023-01-28 | 2023-06-09 | 深圳安侣医学科技有限公司 | Blood imaging analysis system and method |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6049421A (en) * | 1995-07-19 | 2000-04-11 | Morphometrix Technologies Inc. | Automated scanning of microscope slides |
| US6640014B1 (en) * | 1999-01-22 | 2003-10-28 | Jeffrey H. Price | Automatic on-the-fly focusing for continuous image acquisition in high-resolution microscopy |
| US6975400B2 (en) * | 1999-01-25 | 2005-12-13 | Amnis Corporation | Imaging and analyzing parameters of small moving objects such as cells |
| US20060192081A1 (en) * | 2001-07-06 | 2006-08-31 | Palantyr Research, Llc | Imaging system, methodology, and applications employing reciprocal space optical design |
| US20060223165A1 (en) * | 2003-07-18 | 2006-10-05 | Digital Bio Technology | Device for counting cells and method for manufacturing the same |
| US7499166B2 (en) * | 2004-05-20 | 2009-03-03 | The Regents Of The University Of California | Wide field imager for quantitative analysis of microarrays |
Family Cites Families (43)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3879106A (en) * | 1973-04-11 | 1975-04-22 | Pelam Inc | Microscope slide cover slip |
| EP0588972A4 (en) * | 1991-06-13 | 1994-09-14 | Abbott Lab | Optical imaging for positioning and cell counting |
| JPH05133904A (en) * | 1991-07-31 | 1993-05-28 | Suzuki Motor Corp | Optical apparatus for determining aggregation pattern |
| JP2826448B2 (en) * | 1993-09-17 | 1998-11-18 | 株式会社日立製作所 | Flow type particle image analysis method and flow type particle image analysis device |
| US5595710A (en) * | 1995-05-25 | 1997-01-21 | Intelligent Medical Imaging, Inc. | Medical slide holder |
| WO1997004418A1 (en) * | 1995-07-19 | 1997-02-06 | Morphometrix Technologies Inc. | Multi-spectral segmentation for image analysis |
| US5754291A (en) * | 1996-09-19 | 1998-05-19 | Molecular Dynamics, Inc. | Micro-imaging system |
| EP1935983B1 (en) * | 1997-05-05 | 2011-06-22 | ChemoMetec A/S | Method for determination of biological particles in blood |
| EP0921415A1 (en) * | 1997-12-02 | 1999-06-09 | European Community | A method for determining quantitatively the content of fissile material in small size particles |
| US7450229B2 (en) * | 1999-01-25 | 2008-11-11 | Amnis Corporation | Methods for analyzing inter-cellular phenomena |
| US20030026762A1 (en) * | 1999-05-05 | 2003-02-06 | Malmros Mark K. | Bio-spectral imaging system and methods for diagnosing cell disease state |
| JP2001174456A (en) * | 1999-12-20 | 2001-06-29 | Toyobo Co Ltd | Device and method for subclassification of leukocyte |
| US6711283B1 (en) * | 2000-05-03 | 2004-03-23 | Aperio Technologies, Inc. | Fully automatic rapid microscope slide scanner |
| US7546210B2 (en) | 2000-06-08 | 2009-06-09 | The Regents Of The University Of California | Visual-servoing optical microscopy |
| JP3962616B2 (en) * | 2001-04-24 | 2007-08-22 | キヤノン株式会社 | Optical equipment |
| US6977722B2 (en) * | 2001-06-29 | 2005-12-20 | Meso Scale Technologies, Llc. | Assay plates, reader systems and methods for luminescence test measurements |
| US7338168B2 (en) * | 2001-07-06 | 2008-03-04 | Palantyr Research, Llc | Particle analyzing system and methodology |
| US6874775B2 (en) * | 2001-11-16 | 2005-04-05 | Nisca Corporation | Document transfer device and image reading apparatus |
| US7018819B2 (en) * | 2001-11-30 | 2006-03-28 | Cellectricon Ab | Method and apparatus for manipulation of cells and cell-like structures focused electric fields in microfludic systems and use thereof |
| JP2003323600A (en) * | 2002-05-02 | 2003-11-14 | Nisca Corp | Imaging apparatus and focusing method in imaging apparatus |
| AU2003900924A0 (en) * | 2003-02-28 | 2003-03-13 | Medsaic Pty Ltd | Imaging device |
| EP1625226A4 (en) * | 2003-03-27 | 2007-01-10 | Bartron Medical Imaging Llc | System and method for rapidly identifying pathogens, bacteria and abnormal cells |
| US7329537B2 (en) | 2003-04-17 | 2008-02-12 | Nexcelom Bioscience, Llc | Micro-pattern embedded plastic optical film device for cell-based assays |
| EP1651947B1 (en) | 2003-07-19 | 2015-11-04 | NanoEnTek, Inc. | Device for counting micro particles |
| JP4475912B2 (en) * | 2003-10-17 | 2010-06-09 | Ｈｏｙａ株式会社 | Condensing optical system, confocal optical system, and scanning confocal endoscope |
| EP2701194B1 (en) | 2004-06-07 | 2018-08-29 | Fluidigm Corporation | Method and apparatus for imaging a microfluidic device under temperature control |
| US7109481B1 (en) * | 2005-04-28 | 2006-09-19 | Thermo Finnigan Llc | Matrix-assisted laser desorption and ionization (MALDI) sample plate releasably coupled to a sample plate adapter |
| JP2006322765A (en) * | 2005-05-17 | 2006-11-30 | Bisho Junkan Kenkyusho Kk | Blood flow observation device of blood capillary of fingertip |
| JP5028791B2 (en) * | 2005-11-10 | 2012-09-19 | 凸版印刷株式会社 | Imaging device and imaging apparatus |
| US8158801B2 (en) * | 2005-09-26 | 2012-04-17 | Life Technologies Corporation | Violet laser excitable dyes and their method of use |
| JP2007122016A (en) * | 2005-09-30 | 2007-05-17 | Sanyo Electric Co Ltd | Optical material and optical element |
| US7555672B2 (en) * | 2006-01-13 | 2009-06-30 | Agilent Technologies, Inc. | Distributed system and method for error recovery |
| SE530750C2 (en) * | 2006-07-19 | 2008-09-02 | Hemocue Ab | A measuring device, a method and a computer program |
| JP4889437B2 (en) * | 2006-10-16 | 2012-03-07 | オリンパス株式会社 | Weak light imaging device |
| JP4953238B2 (en) * | 2006-11-24 | 2012-06-13 | 学校法人立命館 | Observation device for minute observation object, observation system provided with this observation device, and observation method |
| US7769219B2 (en) * | 2006-12-11 | 2010-08-03 | Cytyc Corporation | Method for assessing image focus quality |
| JP2008250136A (en) * | 2007-03-30 | 2008-10-16 | Fujinon Corp | Imaging lens and imaging apparatus |
| JP2009128267A (en) * | 2007-11-27 | 2009-06-11 | Toyobo Co Ltd | Image processing method |
| JP2009135318A (en) * | 2007-11-30 | 2009-06-18 | Fujifilm Corp | Photoelectric conversion device, imaging device and photosensor |
| JP5466825B2 (en) * | 2008-01-23 | 2014-04-09 | シスメックス株式会社 | Cell image processing system, cell image display system, and cell image display method |
| WO2009107321A1 (en) * | 2008-02-28 | 2009-09-03 | 株式会社ニコン | Microscope apparatus and cell culture apparatus |
| US8570370B2 (en) | 2009-08-31 | 2013-10-29 | Bio-Rad Laboratories, Inc. | Compact automated cell counter |
| US8609363B2 (en) | 2010-11-18 | 2013-12-17 | Bio-Rad Laboratories, Inc. | Viability cell counting by differential light absorption |
-
2010
- 2010-08-27 US US12/869,979 patent/US8570370B2/en active Active
- 2010-08-30 AU AU2010286499A patent/AU2010286499B2/en not_active Ceased
- 2010-08-30 CN CN201080045726.7A patent/CN102576406B/en active Active
- 2010-08-30 EP EP10812708.5A patent/EP2473948B1/en active Active
- 2010-08-30 SG SG2012013009A patent/SG178555A1/en unknown
- 2010-08-30 JP JP2012527068A patent/JP5682974B2/en not_active Expired - Fee Related
- 2010-08-30 CA CA2772376A patent/CA2772376C/en not_active Expired - Fee Related
- 2010-08-30 WO PCT/US2010/047143 patent/WO2011026029A1/en active Application Filing
-
2013
- 2013-09-27 US US14/039,682 patent/US9850517B2/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6049421A (en) * | 1995-07-19 | 2000-04-11 | Morphometrix Technologies Inc. | Automated scanning of microscope slides |
| US6640014B1 (en) * | 1999-01-22 | 2003-10-28 | Jeffrey H. Price | Automatic on-the-fly focusing for continuous image acquisition in high-resolution microscopy |
| US6975400B2 (en) * | 1999-01-25 | 2005-12-13 | Amnis Corporation | Imaging and analyzing parameters of small moving objects such as cells |
| US20060192081A1 (en) * | 2001-07-06 | 2006-08-31 | Palantyr Research, Llc | Imaging system, methodology, and applications employing reciprocal space optical design |
| US20060223165A1 (en) * | 2003-07-18 | 2006-10-05 | Digital Bio Technology | Device for counting cells and method for manufacturing the same |
| US7499166B2 (en) * | 2004-05-20 | 2009-03-03 | The Regents Of The University Of California | Wide field imager for quantitative analysis of microarrays |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105492887A (en) * | 2013-03-15 | 2016-04-13 | 理查德·哈理·特纳 | A system and methods for the in vitro detection of particles and soluble chemical entities in body fluids |
| CN106030343A (en) * | 2013-12-17 | 2016-10-12 | 阿兰蒂克微科学股份有限公司 | Dosimeters including lensless imaging systems |
| CN106030343B (en) * | 2013-12-17 | 2019-11-01 | 阿兰蒂克微科学股份有限公司 | Dosimeter including no lens imaging system |
| CN110914666A (en) * | 2017-05-19 | 2020-03-24 | 兴盛生物科技股份有限公司 | System and method for counting cells |
| US11846579B2 (en) | 2017-05-19 | 2023-12-19 | Thrive Bioscience, Inc. | Systems and methods for counting cells |
| CN114067315A (en) * | 2021-10-23 | 2022-02-18 | 广州市艾贝泰生物科技有限公司 | Cell counting method, cell counting device, computer device, and storage medium |
| CN114067315B (en) * | 2021-10-23 | 2022-11-29 | 广州市艾贝泰生物科技有限公司 | Cell counting method, cell counting device, computer device, and storage medium |
Also Published As
| Publication number | Publication date |
|---|---|
| US20140024107A1 (en) | 2014-01-23 |
| US9850517B2 (en) | 2017-12-26 |
| SG178555A1 (en) | 2012-03-29 |
| WO2011026029A1 (en) | 2011-03-03 |
| US20110211058A1 (en) | 2011-09-01 |
| CN102576406B (en) | 2014-07-23 |
| CA2772376C (en) | 2019-10-22 |
| US8570370B2 (en) | 2013-10-29 |
| EP2473948A4 (en) | 2014-07-09 |
| EP2473948A1 (en) | 2012-07-11 |
| AU2010286499B2 (en) | 2016-05-26 |
| EP2473948B1 (en) | 2019-08-21 |
| AU2010286499A1 (en) | 2012-03-15 |
| CA2772376A1 (en) | 2011-03-03 |
| JP5682974B2 (en) | 2015-03-11 |
| JP2013503351A (en) | 2013-01-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102576406B (en) | Compact automated cell counter | |
| US10921234B2 (en) | Image forming cytometer | |
| AU2018369859A1 (en) | Sample carrier for optical measurements | |
| JP6134422B2 (en) | Stem cell observation method and stem cell observation apparatus | |
| US11385452B2 (en) | Method and apparatus for microscopy | |
| CN110133826A (en) | Information acquisition device | |
| CN109313352A (en) | The analysis based on image of sample | |
| JP2021535373A (en) | Image of surface color and liquid contact angle | |
| CA3160695A1 (en) | Analyzing an analyte disposed within a medium | |
| US10921256B2 (en) | Multi-surface image acquisition system, observation device, observation method, screening method, and stereoscopic reconstruction method of subject | |
| CN108802992A (en) | Sample uniform transillumination based on smart mobile phone illuminates camera arrangement | |
| JP2022541454A (en) | Apparatus for observing living cells or sets of living cells | |
| CN111258050A (en) | Small-volume microscopic light source | |
| EA037486B1 (en) | Imaging system for counting and sizing particles in fluid-filled vessels |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |